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Phosphoproteomic screening identifies physiological substrates of the CDKL5 kinase

Mutations in the gene encoding the protein kinase CDKL5 cause a debilitating neurodevelopmental disease termed CDKL5 disorder. The impact of these mutations on CDKL5 function is poorly understood because the substrates and cellular processes controlled by CDKL5 are unclear. Here, we describe a quant...

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Autores principales: Muñoz, Ivan M, Morgan, Michael E, Peltier, Julien, Weiland, Florian, Gregorczyk, Mateusz, Brown, Fiona CM, Macartney, Thomas, Toth, Rachel, Trost, Matthias, Rouse, John
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6293279/
https://www.ncbi.nlm.nih.gov/pubmed/30266825
http://dx.doi.org/10.15252/embj.201899559
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author Muñoz, Ivan M
Morgan, Michael E
Peltier, Julien
Weiland, Florian
Gregorczyk, Mateusz
Brown, Fiona CM
Macartney, Thomas
Toth, Rachel
Trost, Matthias
Rouse, John
author_facet Muñoz, Ivan M
Morgan, Michael E
Peltier, Julien
Weiland, Florian
Gregorczyk, Mateusz
Brown, Fiona CM
Macartney, Thomas
Toth, Rachel
Trost, Matthias
Rouse, John
author_sort Muñoz, Ivan M
collection PubMed
description Mutations in the gene encoding the protein kinase CDKL5 cause a debilitating neurodevelopmental disease termed CDKL5 disorder. The impact of these mutations on CDKL5 function is poorly understood because the substrates and cellular processes controlled by CDKL5 are unclear. Here, we describe a quantitative phosphoproteomic screening which identified MAP1S, CEP131 and DLG5—regulators of microtubule and centrosome function—as cellular substrates of CDKL5. Antibodies against MAP1S phospho‐Ser(900) and CEP131 phospho‐Ser(35) confirmed CDKL5‐dependent phosphorylation of these targets in human cells. The phospho‐acceptor serine residues in MAP1S, CEP131 and DLG5 lie in the motif RPXSA, although CDKL5 can tolerate residues other than Ala immediately C‐terminal to the phospho‐acceptor serine. We provide insight into the control of CDKL5 activity and show that pathogenic mutations in CDKL5 cause a major reduction in CDKL5 activity in vitro and in cells. These data reveal the first cellular substrates of CDKL5, which may represent important biomarkers in the diagnosis and treatment of CDKL5 disorder, and illuminate the functions of this poorly characterized kinase.
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spelling pubmed-62932792018-12-18 Phosphoproteomic screening identifies physiological substrates of the CDKL5 kinase Muñoz, Ivan M Morgan, Michael E Peltier, Julien Weiland, Florian Gregorczyk, Mateusz Brown, Fiona CM Macartney, Thomas Toth, Rachel Trost, Matthias Rouse, John EMBO J Resource Mutations in the gene encoding the protein kinase CDKL5 cause a debilitating neurodevelopmental disease termed CDKL5 disorder. The impact of these mutations on CDKL5 function is poorly understood because the substrates and cellular processes controlled by CDKL5 are unclear. Here, we describe a quantitative phosphoproteomic screening which identified MAP1S, CEP131 and DLG5—regulators of microtubule and centrosome function—as cellular substrates of CDKL5. Antibodies against MAP1S phospho‐Ser(900) and CEP131 phospho‐Ser(35) confirmed CDKL5‐dependent phosphorylation of these targets in human cells. The phospho‐acceptor serine residues in MAP1S, CEP131 and DLG5 lie in the motif RPXSA, although CDKL5 can tolerate residues other than Ala immediately C‐terminal to the phospho‐acceptor serine. We provide insight into the control of CDKL5 activity and show that pathogenic mutations in CDKL5 cause a major reduction in CDKL5 activity in vitro and in cells. These data reveal the first cellular substrates of CDKL5, which may represent important biomarkers in the diagnosis and treatment of CDKL5 disorder, and illuminate the functions of this poorly characterized kinase. John Wiley and Sons Inc. 2018-09-28 2018-12-14 /pmc/articles/PMC6293279/ /pubmed/30266825 http://dx.doi.org/10.15252/embj.201899559 Text en © 2018 The Authors. Published under the terms of the CC BY 4.0 license This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Resource
Muñoz, Ivan M
Morgan, Michael E
Peltier, Julien
Weiland, Florian
Gregorczyk, Mateusz
Brown, Fiona CM
Macartney, Thomas
Toth, Rachel
Trost, Matthias
Rouse, John
Phosphoproteomic screening identifies physiological substrates of the CDKL5 kinase
title Phosphoproteomic screening identifies physiological substrates of the CDKL5 kinase
title_full Phosphoproteomic screening identifies physiological substrates of the CDKL5 kinase
title_fullStr Phosphoproteomic screening identifies physiological substrates of the CDKL5 kinase
title_full_unstemmed Phosphoproteomic screening identifies physiological substrates of the CDKL5 kinase
title_short Phosphoproteomic screening identifies physiological substrates of the CDKL5 kinase
title_sort phosphoproteomic screening identifies physiological substrates of the cdkl5 kinase
topic Resource
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6293279/
https://www.ncbi.nlm.nih.gov/pubmed/30266825
http://dx.doi.org/10.15252/embj.201899559
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